Treatment of Chronic Experimental Autoimmune Encephalomyelitis with Epigallocatechin-3-Gallate and Glatiramer Acetate Alters Expression of Heme-Oxygenase-1.

We previously demonstrated that epigallocatechin-3-gallate (EGCG) synergizes with the immunomodulatory agent glatiramer acetate (GA) in eliciting anti-inflammatory and neuroprotective effects in the relapsing-remitting EAE model. Thus, we hypothesized that mice with chronic EAE may also benefit from this combination therapy. We first assessed how a treatment with a single dose of GA together with daily application of EGCG may modulate EAE. Although single therapies with a suboptimal dose of GA or EGCG led to disease amelioration and reduced CNS inflammation, the combination therapy had no effects. While EGCG appeared to preserve axons and myelin, the single GA dose did not improve axonal damage or demyelination. Interestingly, the neuroprotective effect of EGCG was abolished when GA was applied in combination. To elucidate how a single dose of GA may interfere with EGCG, we focused on the anti-inflammatory, iron chelating and anti-oxidant properties of EGCG. Surprisingly, we observed that while EGCG induced a downregulation of the gene expression of heme oxygenase-1 (HO-1) in affected CNS areas, the combined therapy of GA+EGCG seems to promote an increased HO-1 expression. These data suggest that upregulation of HO-1 may contribute to diminish the neuroprotective benefits of EGCG alone in this EAE model. Altogether, our data indicate that neuroprotection by EGCG in chronic EAE may involve regulation of oxidative processes, including downmodulation of HO-1. Further investigation of the re-dox balance in chronic neuroinflammation and in particular functional studies on HO-1 are warranted to understand its role in disease progression

GA does not influence the anti-inflammatory effects of EGCG-treatment <i>in vivo</i>.

<p>To elucidate mechanisms of action of EGCG, mice with a minimum score of 0.5 were treated for 12 days with EGCG alone or with combination of EGCG and GA, starting at day 14 after immunization. A: Disease severity of EGCG and combination therapy group. Analysis includes data from two independent experiments (Mann-Whitney). B: Proliferation of MOG-specific CD4⁺ T-cells day 26 after immunization. C: Mononuclear cells from CNS were stimulated with anti-CD3 and anti-CD28. Cells were stained with anti-CD4, anti-CD45 as surface markers and with anti-IFNγ, anti-FoxP3 and anti-IL17. d) RNA extracted from the CNS tissue (brains and spinal cords) was used to determine T-bet, RORγt, IFNγ and IL17 mRNA expression using quantitative real-time RT-PCR. (t-test) *p<0.05.</p

The application of GA in combination with EGCG interferes with the anti-inflammatory and neuroprotective effects of EGCG single therapy.

<p>A: Quantification of inflammation, B: neurodegeneration C: and degree of demyelination in the spinal cord from control, GA, EGCG and combination therapy group included in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0130251#pone.0130251.g001" target="_blank">Fig 1</a>. Histopathological changes were assessed semi-quantitatively as percentage of spinal cord quadrants that showed pathological changes related to all investigated tissue quadrants. Representative Hematoxylin and Eosin staining to monitor inflammation of transverse spinal cord sections of control (D), GA treated mice (G), EGCG treated mice (J) and mice treated with the combination therapy (M) are shown. Bielschowsky staining was used to assess axonal damage. Representative spinal cord sections of control (E), GA treated mice (H), EGCG treated mice (K) and mice treated with the combination therapy (N) are shown. The degree of demyelination was determined by Luxol Fast Blue staining. Representative LFB spinal cord sections of control group (F), GA treated group (I), EGCG treated group (L) and group treated with EGCG and GA (O) are shown. (Kruskal-Wallis) *p<0.05, **p<0.01.</p

Treatment onset and mean maximum clinical score.

<p>Treatment onset and mean maximum clinical score.</p

Industrie 4.0 - Interoperabilität durch OPC UA mit Companion Specifications : Mehrwerte für Stakeholder des Maschinen- und Anlagenbaus

Interoperabilität ist eine der wichtigsten Säulen der digitalen Transformation und Teil der Vision 2030 der Plattform Industrie 4.0. Erfolgsentscheidend für die digitale Transformation eines jeden Unternehmens ist jedoch nicht der technologische Reifegrad der Digitalisierung, sondern die zielgerichtete Nutzung bestehender Digitalisierungslösungen, -konzepte und hierfür relevante Standards. Letztere sollten bewusst gewählt und möglichst flächendeckend genutzt werden. Die entscheidende Frage ist in diesem Zusammenhang, welche Mehrwerte sich durch den Einsatz der jeweiligen Standards realisieren lassen. Es gilt, sowohl auf Herstellerseite als auch Betreiberseite, relevante Anwendungsfälle durch Nutzung von Standards effizienter, skalierbarer, sicher und letztendlich schneller bzw. somit kostenoptimaler umzusetzen. Eine für den Maschinen- und Anlagenbau, sowie seine Kundenindustrien, im Fokus stehende Interoperabilitätslösung sind standardisierte Schnittstellen auf Basis der OPC UA Technologie. Dazu gehören, neben der sicheren von der Produktion bis in die Cloud skalierbaren Kommunikation über OPC UA, auch die sogenannte OPC UA Companion Specifications (CS) für die präzise Standardisierung von Produktionsinformationen und den zugehörigen Metadaten. Die Weltsprache der Produktion bestehend aus Grammatik (OPC UA) und Vokabular (OPC UA CS) ist bereits heute real verfügbar und wächst rasant. Die OPC UA for Machinery mit ihren zahlreichen Building Blocks ist eine zentrale Grundlage für die Entwicklung der vielen domänenspezifischen Companion Specifications. Der Leitfaden „Industrie 4.0 Interoperabilität durch OPC UA mit Companion Specifications — Mehrwerte für die Stakeholder des Maschinen- und Anlagenbau“ schließt die Lücke zwischen den verfügbaren sowie in Erarbeitung befindlichen Entwicklungen und den realisierbaren Szenarien für die diskrete und kontinuierliche Fertigung. Es werden für die einzelnen Rollen in Unternehmen konkrete Vorteile und Mehrwerte aufgezeigt. Als ein Leitfaden zum Thema Interoperabilität bauen die Inhalte auf der Neuauflage des Leitfaden „Industrie 4.0 Kommunikation mit OPC UA — Leitfaden zur Einführung in den Mittelstand“ von 2023 auf und geben somit ein umfassendes Gesamtbild, wie Interoperabilität im Maschinen- und Anlagenbau mehrwertbringend eingesetzt werden kann. Experten aus der Industrie und Forschung haben die zentralen Anwendungsfälle Asset Management, Machine & Component Monitoring, KPI Berechnungen wie OEE, Job sowie Energy Management identifiziert und veranschaulichen den konkreten Einsatz von OPC UA mit Companion Specifications aus Sicht der herstellenden und betreibenden Unternehmen

Effects of EGCG alone and in combination with a suboptimal dose of GA in established EAE.

<p>A: Disease severity of control, GA, EGCG and combination therapy group. Analysis includes data from three independent experiments (Mann-Whitney test). B: Mean clinical scores of animals are shown. Data are given as mean ± SEM. Cumulative disease activity is represented as the area under the curve of control, GA, EGCG and combination therapy group (Kruskal-Wallis). C: Proliferation of MOG-specific CD4⁺ T cells at day 62 after immunization. CFSE-labeled lymph node cells were incubated for 72h with MOG (50μg/ml). As a positive control, cells were cultured with 3 μg/ml anti-CD3 antibody and 2.5 μg/ml anti-CD28 antibody. For the negative control, cells were incubated alone, in the absence of antigen. To assess cell division cells were stained with anti-CD4 Alexa Fluor 647 and analyzed by flow cytometry (ANOVA). *p<0.05, **p<0.01, ***p<0.001.</p

Addition EGCG therapy leads to decreased expression of heme oxygenase-1 (HO-1).

<p>A: Relative mRNA expression of HO-1 in cerebral and cerebellar regions as well as spinal cords of mice treated with vehicle control, EGCG alone, or in combination with GA. (ANOVA). B: Relative expression of HO-1 in mice treated with EGCG compared to mice treated with EGCG+GA C: Relative expression of HO-1 in mice treated with EGCG compared to control vehicle treated animals (t-test).</p

The combined application of GA and EGCG treatment does not alter the iron chelator activity but enhanced HO-1 expression compared to EGCG.

<p>A: Soluble iron content in blood serum was quantified day 26 after immunization by using a modification of the ferrozine-based assay. B: Relative mRNA expression of HO-1 in cerebellar and cerebral regions of the CNS of mice included in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0130251#pone.0130251.g003" target="_blank">Fig 3</a> treated with EGCG alone, or in combination with GA. (t-test) *p<0.05.</p

Thermal (n,γn,{\gamma}) cross section and resonance integral of 171Tm^{171}\mathrm{Tm}}

International audienceBackground: About 50% of the heavy elements are produced in stars during the slow neutron capture process. The analysis of branching points allows us to set constraints on the temperature and the neutron density in the interior of stars. Purpose: The temperature dependence of the branch point Tm171 is weak. Hence, the Tm171 neutron capture cross section can be used to constrain the neutron density during the main component of the s process in thermally pulsing asymptotic giant branch (TP-AGB) stars. Methods: A Tm171 sample produced at the ILL was activated with thermal and epithermal neutrons at the TRIGA research reactor at the Johannes Gutenberg-Universität Mainz. Results: The thermal neutron capture cross section and the resonance integral have been measured for the first time to be σth=9.9±0.9b and σRI=193±14b. Conclusions: Based on our results, new estimations of the direct capture components' impact on the Maxwellian-nAveraged cross sections (MACS) are possible

Critical Role for Asparagine Endopeptidase in Endocytic Toll-like Receptor Signaling in Dendritic Cells

SummaryIntracellular Toll-like receptor 3 (TLR3), TLR7, and TLR9 localize in endosomes and recognize single-stranded RNA and nucleotides from viruses and bacteria. This interaction induces their conformational changes resulting in the production of proinflammatory cytokines and upregulation of cell surface molecules. TLR9 requires a proteolytic cleavage for its signaling. Here, we report that myeloid and plasmacytoid dendritic cells (DCs) deficient for the asparagine endopeptidase (AEP), a cysteine lysosomal protease, showed a decrease in the secretion of proinflammatory cytokines in response to TLR9 stimulation in vitro and in vivo. Upon stimulation, full-length TLR9 was cleaved into a 72 kDa fragment and this processing was strongly reduced in DCs lacking AEP. Processed TLR9 coeluted with the adaptor molecule MyD88 and AEP after size exclusion chromatography. When expressed in AEP-deficient DCs, the 72 kDa proteolytic fragment restored TLR9 signaling. Thus, our results identify an endocytic protease playing a critical role in TLR processing and signaling in DCs